Type I Interferon Response Is Delayed in Human Astrovirus Infections

Type I interferon (IFN) activation and its subsequent effects are important in the response to viral infections. Here we show that human astroviruses (HAstVs), which are important agents of acute gastroenteritis in children, induce a mild and delayed IFN response upon infecting CaCo-2 cells. Although IFN-β mRNA is detected within infected cells and supernatant from infected cells show antiviral activity against the replication of other well-known IFN-sensitive viruses, these responses occur at late stages of infection once genome replication has taken place. On the other hand, HAstV replication can be partially reduced by the addition of exogenous IFN, and inhibition of IFN activation by BX795 enhances viral replication, indicating that HAstVs are IFN-sensitive viruses. Finally, different levels of IFN response were observed in cells infected with different HAstV mutants with changes in the hypervariable region of nsP1a/4, suggesting that nsP1a/4 genotype may potentially have clinical implications due to its correlation with the viral replication phenotype and the antiviral responses induced within infected cells.     

Human astrovirus coat protein inhibits serum complement activation via C1, the first component of the classical pathway

Human astroviruses (HAstVs) belong to a family of nonenveloped, icosahedral RNA viruses that cause noninflammatory gastroenteritis, predominantly in infants. Eight HAstV serotypes have been identified, with a worldwide distribution. While the HAstVs represent a significant public health concern, very little is known about the pathogenesis of and host immune response to these viruses. Here we demonstrate that HAstV type 1 (HAstV-1) virions, specifically the viral coat protein (CP), suppress the complement system, a fundamental component of the innate immune response in vertebrates. HAstV-1 virions and purified CP both suppress hemolytic complement activity. Hemolytic assays utilizing sera depleted of individual complement factors as well as adding back purified factors demonstrated that HAstV CP suppresses classical pathway activation at the first component, C1. HAstV-1 CP bound the A chain of C1q and inhibited serum complement activation, resulting in decreased C4b, iC3b, and terminal C5b-9 formation. Inhibition of complement activation was also demonstrated for HAstV serotypes 2 to 4, suggesting that this phenomenon is a general feature of these human pathogens. Since complement is a major contributor to the initiation and amplification of inflammation, the observed CP-mediated inhibition of complement activity may contribute to the lack of inflammation associated with astrovirus-induced gastroenteritis. Although diverse mechanisms of inhibition of complement activation have been described for many enveloped animal viruses, this is the first report of a nonenveloped icosahedral virus CP inhibiting classical pathway activation at C1.     

Astrovirus MLB1 is not associated with diarrhea in a cohort of Indian children

Astroviruses are a known cause of human diarrhea. Recently the highly divergent astrovirus MLB1 (MLB1) was identified in a stool sample from a patient with diarrhea. It has subsequently been detected in stool from individuals with and without diarrhea. To determine whether MLB1 is associated with diarrhea, we conducted a case control study of MLB1. In parallel, the prevalence of the classic human astroviruses (HAstVs) was also determined in the same case control cohort. 400 cases and 400 paired controls from a longitudinal birth cohort in Vellore, India were analyzed by RT-PCR. While HAstVs were associated with diarrhea (p = 0.029) in this cohort, MLB1 was not; 14 of the controls and 4 cases were positive for MLB1. Furthermore, MLB1 viral load did not differ significantly between the cases and controls. The role of MLB1 in human health still remains unknown and future studies are needed.     

Molecular Evolution of GII-4 Norovirus Strains

BACKGROUND: Human Noroviruses (NoV) are the major cause of acute nonbacterial gastroenteritis and the leading cause of outbreaks of gastroenteritis worldwide. Genotype II-4 (GII-4) NoV has been shown to spread rapidly and is the most commonly detected strain worldwide, particularly in association with outbreaks. Previously, we have shown that circulating GII-4 NoV strains exist as populations of selectively neutral variants, and that the emergence of epidemic GII-4 NoV strains correlated with mutations in at least two key sites (Sites A and B) within the P2 domain of the surface exposed major capsid protein (VP1). METHODOLOGY: We developed a rapid pyrosequencing method for screening of the two Sites A and B and a homology based modelling system was used to predict the effects of amino acid substitutions at these sites on the antigenic properties of the virus (defined as surface motif types). PRINCIPLE FINDING/CONCLUSION: Here, we describe the characterisation of amino acid diversity at Sites A and B for 1062 GII-4 NoV strains from clinical specimen associated with outbreak of gastroenteritis (2000-2011) and 250 GII-4 NoV sequences from Genbank. Our data identified a high diversity of different Site A and B site combinations at amino acid level and amino acid diversity was higher at Site B than Site A. Site A motifs could be grouped into 3 clusters based on similar surface motif types. We predict that Site A is a major epitope on the virus surface, responsible for defining the antigenic profile, and a more subtle role for Site B, maintaining minor antigenic variation within the virus population.     

Human Bocaviruses Are Not Significantly Associated with Gastroenteritis: Results of Retesting Archive DNA from a Case Control Study in the UK

Gastroenteritis is a common illness causing considerable morbidity and mortality worldwide. Despite improvements in detection methods, a significant diagnostic gap still remains. Human bocavirus (HBoV)s, which are associated with respiratory infections, have also frequently been detected in stool samples in cases of gastroenteritis, and a tentative association between HBoVs, and in particular type-2 HBoVs, and gastroenteritis has previously been made. The aim of this study was to determine the role of HBoVs in gastroenteritis, using archived DNA samples from the case-control Infectious Intestinal Disease Study (IID). DNA extracted from stool samples from 2,256 cases and 2,124 controls were tested for the presence of HBoV DNA. All samples were screened in a real time PCR pan-HBoV assay, and positive samples were then tested in genotype 1 to 3-specific assays. HBoV was detected in 7.4% but no significantly different prevalence was observed between cases and controls. In the genotype-specific assays 106 of the 324 HBoV-positive samples were genotyped, with HBoV-1 predominantly found in controls whilst HBoV-2 was more frequently associated with cases of gastroenteritis (p<0.01). A significant proportion of HBoV positives could not be typed using the type specific assays, 67% of the total positives, and this was most likely due to low viral loads being present in the samples. However, the distribution of the untyped HBoV strains was no different between cases and controls. In conclusion, HBoVs, including HBoV-2 do not appear to be a significant cause of gastroenteritis in the UK population.     

Full sequence analysis and characterization of a human astrovirus type 1 isolate from South Korea

Human astroviruses are recognized as an important cause of infantile gastroenteritis around the world. In South Korea, sporadic cases of HAstV infection have been reported since 2002. However, hitherto, there have been no studies reporting the whole genome sequence of an HAstV isolate from South Korea. Hence, we sequenced and analyzed the entire genome of an HAstV-1 strain (lhar) that was isolated in Seoul, South Korea. The whole-genome sequence analysis revealed 3 open reading frames comprising the whole genome: ORF1a (2,763 bp), ORF1b (1,548 bp), and ORF2 (2,364 bp). The lhar strain showed amino acid identities with 8 other reference strains of 87.6–98.7%, 94.2–98.8%, and 62.6–99.0% in the ORF1a, ORF1b, and ORF2 regions, respectively. The amino acid sequence of the capsid region encoded by ORF2 was compared with a total of 19 HAstV-1 strains and 8 HAstVs reference strains isolated in various countries. This revealed 1 amino acid substitution, at aa412 (Pro → Arg) in ORF2. This study, the first to report the full-length sequence of an HAstV isolated in South Korea, is meaningful in that it can be used as a full-length HAstV sequence standard for future comparison studies. It may also prove useful to the field of public health field by facilitating the diagnosis and the prediction of new emerging variants.     

The past and future of farmland birds in Hungary

Capsule Populations of birds on farmland are larger and more stable in Hungary than in the UK and may provide baseline targets when planning population restoration programmes in more intensively farmed regions of Europe.

Aims To review the available evidence on farmland bird populations and their changes over the past century in Hungary, and to compare this with similar data for the UK.

Methods Published papers and grey literature were searched to determine long-term bird population trends for birds on farmland in Hungary, and for research evidence on the relationship between farmland management and bird diversity in Hungary.

Results Population density of common farmland birds is higher and trends are more positive in Hungary compared to the UK. These findings correlate with the recent change to generally less intensive agriculture in Hungary. However, while the birdlife associated with farmland in Hungary can be considered to have high diversity and density, it is still lower than it was in the first half of the 20th century and earlier. The few studies available showed that low-intensity traditional management promotes a rich biodiversity in both grasslands and arable systems in Hungary. Agri-environment schemes were introduced when Hungary joined the EU in 2004; however, their influence on biodiversity has not been systematically monitored.

Conclusions Insights emerging from farmland bird research in those European countries which still practice extensive agricultural techniques could be used to set general baseline targets for restoring biodiversity in regions where farmlands are now intensively managed. At the European scale urgent tasks are to: (1) investigate the relationships between management and bird diversity and density on a much wider geographical scale, (2) evaluate the geographical generality of the existing evidence base (which is mainly based on studies conducted in more intensively farmed regions), and (3) enhance the policy impact of conservation research.     

Detection and molecular characterization of adenoviruses in Korean children hospitalized with acute gastroenteritis

Human adenoviruses (HAdVs) are an important cause of acute gastroenteritis in children. However, few studies on the epidemiology or types of HAdVs associated with acute gastroenteritis have been conducted in Korea. Therefore, in the present study, the incidence of HAdV in 2064 stool samples from Korean children hospitalized with acute gastroenteritis (2004-2006) was assessed and the types of viruses present determined. Polymerase chain reaction, sequencing, and phylogenic analyses revealed that 113 samples (5.5%) were HAdV-positive. While HAdVs were mainly detected during July to October, no seasonal difference between the enteric and non-enteric viruses in the incidence of HAdV was observed. HAdV-41 and HAdV-40 were found in 54 (47.8%) and 3 (2.6%) HAdV-positive samples, respectively. HAdV-3, HAdV-7, HAdV-2, HAdV-31, HAdV-4, and HAdV-37 were detected in 11 (9.7%), 5 (4.4%), 2 (1.7%), 2 (1.7%), 1 (0.8%), and 1 (0.8%) of sample(s), respectively. Thus, not only enteric, but also non-enteric, HAdVs may play an important role in acute gastroenteritis in Korean children.     

Assessing stream dwelling caddisfly assemblages (Insecta: Trichoptera) collected by light traps in Hungary

In the present study the conservation status of light trap-collected stream dwelling caddisfly assemblages was evaluated on the basis of original data and also from the literature representing different regions in Hungary. Altogether the catches of eight complete seasons were compared. Species richness, diversity and endangerment indices as well as a newly introduced rarity index (RI) expressing the average rarity of the collected species in Hungary were calculated to evaluate the conservation status of the streams in the Bakony, Börzsöny and Bükk Mountains and in the Zala Hills. The results highlight the importance of streams in maintaining endangered species of caddisfly in Hungary and bring to attention the fact that species richness or diversity are not necessarily the best indicators of conservation status of caddisfly assemblages. Indicators, for instance RI, which take into consideration the general rarity of the species seem to reflect more sharply conservation status and thus are more appropriate for the assessment of caddisfly assemblages.     

A food‐borne outbreak of gastroenteritis due to genotype G1P[8] rotavirus among adolescents in Japan

Six high school students in Tochigi prefecture, Japan, developed gastroenteritis after eating at a pork cutlet shop. Molecular epidemiologic analyses showed that the causative agent was genotype G1P[8] rotavirus (RV), this being detected in stool samples from both the patients and the asymptomatic food handlers. The detected RV strains were closely related genetically. The only uncooked food that all victims had eaten was raw sliced cabbage. These findings results suggest that uncooked foods contaminated with RV may be sources of infectious gastroenteritis in adolescents.     

Spatial distribution and diversity of chironomid assemblages in a small hilly stream (Diptera: Chironomidae)

In 2006–2007, spatial variations of chironomid assemblages of a small chalky watercourse were investigated in the hilly region of Balaton Upland, Hungary. Samplings were carried out every three weeks at seven characteristically different sites along the whole length of the watercourse. Altogether 2966 specimens were collected and identified. The chironomid fauna of the stream was relatively rich with 40 taxa belonging to four subfamilies (eight Tanypodinae, one Prodiamesinae, 17 Orthocladiinae, 14 Chironominae). The lowest number of species and number of individuals was found at the spring, and the highest at the middle reach. Based on multivariate analysis the different sections of the creek were characterised by distinctive chironomid communities. Although longitudinal changes in the chironomid community of the Örvényesi creek were detectable due to changes in natural environmental factors, land-use changes also significantly influenced the chironomid community composition and diversity. The natural and anthropogenic effects were hardly distinguishable. Furthermore, we found that the response of different diversity indices to environmental variability was different. This draws attention to the difficulties in evaluating the relationships between stream communities and environmental variables using different diversity indices. The taxonomic diversity, which incorporates the taxonomical relationships between the specimens, proved to be one of the most useful diversity indices to characterise the stream chironomid assemblages.     

Molecular epidemiology of adenovirus infection among pediatric population with diarrhea in Asia

A total of 3,577 fecal specimens from infants and children with acute gastroenteritis in Japan, Korea and Vietnam during 1998 and 2001 were tested for adenovirus by the ELISA method. Of these, adenovirus was detected in 158 (4.4%). The detection rate of adenovirus was highest in Korea (8.7%, 20/231) followed by 5.0% (100/1,991) in Japan and 2.8% (38/1,355) in Vietnam. All adenoviruses were further serotyped by PCR-RFLP. The diversity of adenovirus serotypes including Ad2, 3, 5, 8, 31, 40 and 41 was demonstrated. Worth of note was a decrease in the rate of isolation of Ad40 (7.6%, 12/158) and a concomitant increase of Ad41 (63.9%, 101/158) to become the predominant serotype. Another interesting feature of the study was the presence of Ad3 (4.0% and 30%) and Ad8 (8.0% and 20%) in Japan and Korea, respectively, which is mainly associated with keratoconjunctivitis worldwide. Our result underscored the importance of adenovirus in association with acute gastroenteritis in Asian countries.     

Molecular characterization of rotaviruses obtained from patients with rotavirus-associated encephalitis/encephalopathy

Group A rotavirus (RVA) rarely causes severe complications such as encephalitis/encephalopathy. However, the pathophysiology of this specific complication remains unclear. Next-generation sequence analysis was used to compare the entire genome sequences of RVAs detected in patients with encephalitis/encephalopathy and gastroenteritis. This study enrolled eight patients with RVA encephalitis/encephalopathy and 10 with RVA gastroenteritis who were treated between February 2013 and July 2014. Viral RNAs were extracted from patients' stool, and whole-genome sequencing analysis was carried out to identify the specific gene mutations in RVA obtained from patients with severe neurological complications. Among the eight encephalitis/encephalopathy cases, six strains were DS-1-like G1P[8] and the remaining two were Wa-like G1P[8] (G1-P[8]-I1-R1-C1-M1-A1-N1-T1-E1-H1). Meanwhile, eight of the 10 viruses detected in rotavirus gastroenteritis patients were DS-1-like G1P[8], and the remaining two were Wa-like G1P[8]. These strains were further characterized by conducting phylogenetic analysis. No specific clustering was demonstrated in RVAs detected from encephalitis/encephalopathy patients. Although the DS-1-like G1P[8] strain was predominant in both groups, no specific molecular characteristics were detected in RVAs from patients with severe central nervous system complications.     

The air spora of an orchid greenhouse

The orchid collection of the ELTE Botanical Garden, Budapest, Hungary was monitored for airborne fungi using viable and non-viable air-sampling methods (Hirst-type and a 3-stage Andersen sampler) with three different culture media. A new culture method was also applied to identify fungal spores from Hirst-type samples. The aim of this study was to determine the diversity, human- and phytopathological potential of the air spora. To find out sources of airborne fungi, samples were collected from the air in an adjacent greenhouse and outdoors, and from necrotic plants. A total of 58 genera were found in the air samples. Cladosporium and Penicillium spp. were common members of the airborne biota. A high proportion (27.5%) of identified genera may be presented as a member of microbial consortium associated with the orchids. Airborne fungi potentially pathogenic to humans were also detected. One species, Zygosporium masonii, was new to Hungary. Statistical analysis indicated that conditions of sampling had significant effects. The principal component analysis elucidated the three principal components representing 75.34% of the total variance; the clusters of variables were related to the three types of culture media. Relative abundance of small-sized spores was high, presumably because of the fungal species composition and accelerated sedimentation of large spores in still air. Apparently, in the studied orchid greenhouse, a specific mycobiota developed due to the climate and hosts (Orchideaceae) grown there.